Cell/tissue culture apparatus

ABSTRACT

A cell/tissue culture apparatus is suitable for a culture of a cultivated object of a cell or tissue adapted for a prescribed portion of a living body of a breast and so on, a culture chamber ( 18 , a culture vessel  10 ) which accommodates a cell or tissue to be cultivated and also makes a culture fluid ( 34 ) circulate is formed by a flexible material (flexible films  12, 14 ), and the culture chamber is immersed in a fluid (water  8 ). A pressure or an oscillation is given to the culture chamber installed in the fluid, and a physical stimulation due to the pressure from the fluid or the oscillation of the fluid is given to a cell or tissue under a culture.

TECHNICAL FIELD

The present invention relates to a cell/tissue culture apparatus usedfor a culture of a cell or tissue, and so on, to which tissueengineering is applied. More particularly, the present invention relatesto a cell/tissue culture apparatus for efficiently realizing ametabolism function of a cell or tissue when performing an in vitroculture of a cell or tissue of a living body of a human body and so on,and for giving a physical stimulation necessary for prolongation,differentiation and promotion of a cell to an cultivated object.

BACKGROUND ART

In a conventional method which performs an in vitro culture of a cell ortissue of a living body of a human body and so on, there is employed amethod in which temperature, humidity, the concentration of carbondioxide and the concentration of oxygen in an incubator (a culturehouse) are maintained at proper conditions, and a cell is cultivated inthat incubator. A cell or tissue is placed in a culture fluid in afloating state, or a cell or tissue is fixed to an inside or a surfaceof a gel in which an ingredient of the culture fluid is input, andthereby, the cell or tissue is made to proliferate and to grow. Or, acell or tissue is transplanted in a material, which is called a matrixor a scaffold, a carrier or a mold, and so on (herein after called“matrix” simply), and thereby, the cell or tissue is made to proliferateand to grow.

In this case, for the proliferation and growth of the cell or tissue, itis important to give a physical stimulation to the cell or tissue to becultivated in addition to environmental conditions of temperature,humidity, the concentration of carbon dioxide, the concentration ofoxygen, and so on. Such a physical stimulation is an indispensablefactor for promoting differentiation and growth of a cell or tissue andfor growing to a cell or tissue closer to a cell or tissue in a livingbody. As a technology which gives a physical stimulation to theproliferation and growth of a cell or tissue, there are, for example,the Japanese Official Announcement Patent Publication No.2001-504697entitled “APPLICATION OF SHEAR FLOW STRESS TO CHONDROCYTES” and theJapanese Patent No.3163533 entitled “LOADING DEVICE OF EXPANDING ANDCONTRACTING STIMULATION FOR CULTURING CELL BY USING SILICON BELT”.

By the way, as for a restoring method of a breast to a patient whom aneliminating operation of the breast has been performed owing to breastcancer and soon, a method which injects silicone into an excisedportion, and so on are general. The silicone as a material for restoringa shape is much used as a material adapted to a living body, which isnon-absorbability toward a living body. However, if it is maintained fora long term in a living body, there is indication that an outflow of thesilicone into the living body can not be avoided, and so on. Because ofthis, without using a foreign substance of the silicone and so on,restoration of a deficient portion by means of a tissue similar to aliving body is strongly desired.

The present invention, therefore, makes it an object to provide acell/tissue culture apparatus suitable for a culture of a cultivatedobject of a cell or tissue which is adapted to a prescribed portion of aliving body of a breast and so on.

DISCLOSURE OF THE INVENTION

A cell/tissue culture apparatus of the present invention makes it afeature to form a culture chamber (18, a culture vessel 10), whichaccommodates a cell or tissue to be cultivated and also makes a culturefluid (34) circulate, by a flexible material (flexible films 12, 14),and to immerse this culture chamber in a fluid (water 8). If the cell ortissue to be cultivated is housed in the culture chamber formed by theflexible material, if the culture fluid is made to circulate, and, alongwith these, if the culture chamber is made to immerse in the fluid, itis possible to apply a pressure from the fluid to an inside of theculture chamber from an outside of the culture chamber, and thispressure becomes a physical stimulation toward the cell or tissue to becultivated. Therefore, it is possible to cultivate a tissue bodyaccording to a shape of a desired portion in a human body, and it isalso possible to realize flexibility and toughness which the portionhas.

In the cell/tissue culture apparatus of the present invention, theapparatus makes it a feature to give a physical stimulation to said cellor tissue in said culture camber by making said fluid oscillate. If thecell or tissue to be cultivated is accommodated in the culture chamberformed by flexible material, if the culture fluid is made to circulate,if the culture chamber is made to immerse in the fluid, and, along withthese, if a pressure and/or a vibration is applied to an inside of theculture chamber with the fluid used as a medium from an outside of theculture chamber, an oscillation of the fluid is applied to the cell ortissue in the culture chamber as a physical stimulation.

In the cell/tissue culture apparatus of the present invention, saidculture chamber makes it a feature to provide a space part of a shapeaccording to a shape of cells or tissue to be cultivated. That is, in acell or tissue to be cultivated, for example, there are cases in whichit is rendered necessary to cultivate in a shape of a portion of a humanbody to be restored. Because of this, if the space part of the culturechamber corresponds with a shape of said portion, it is possible to makethe culture chamber function as a mold for formation, and it is possibleto cultivate/produce a tissue body corresponding to the shape of adesired portion.

In the cell/tissue culture apparatus of the present invention, saidoscillation of the fluid makes it a feature to be due to a jet of fluid.That is, the jet of fluid can generate an optional oscillation in thefluid. If this jet is applied through the fluid to the culture chamber,it is possible to give a physical stimulation of pressure, vibration andsoon according to modes of strength and so on of the jet to a cell ortissue to be cultivated. This physical stimulation can be controlledaccording to modes of strength, direction and so on of the jet.

In the cell/tissue culture apparatus of the present invention, saidoscillation of the fluid makes it a feature to be due to wave motion.That is, there are a transverse wave and a longitudinal wave in the wavemotion, and the wave motion has variational factors of a period, a leveland so on. Hence, if an oscillation is generated in the fluid by thiswave motion, control of the oscillation is easy by means of anadjustment of the wave motion, and it is possible to generate wavemotion which is desired. As a result, it is possible to give a physicalstimulation, which is desired, to a cell or tissue.

In the cell/tissue culture apparatus of the present invention, saidphysical stimulation makes it a feature to be a shearing stress. Bygiving the shearing stress to a cell or tissue, a tough tissue can beobtained.

In the cell/tissue culture apparatus of the present invention, saidphysical stimulation makes it a feature to be given intermittently orcontinuously. In order to cultivate a tissue equivalent to a human body,by making intermittence or continuousness of the physical stimulation,it is possible to give a stimulation equivalent to a human body to acell or tissue.

And, the objects, features, advantages and so on of the presentinvention will be more clear by referring to an explanation in the modesfor carrying out the present invention and embodiments shown in thedrawings.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a drawing showing a first embodiment of a cell/tissue cultureapparatus according to the present invention.

FIG. 2 is a perspective view showing a culture unit.

FIG. 3 is a block diagram showing an embodiment of a culture controldevice.

FIG. 4 is a flow chart showing a control program.

FIG. 5 is a flow chart showing a control program continued on FIG. 4.

FIG. 6 is a flow chart showing a control program continued on FIG. 5.

FIG. 7 is a timing chart showing a physical stimulation.

FIG. 8 is a drawing showing a second embodiment of a cell/tissue cultureapparatus according to the present invention.

BEST MODES FOR CARRYING OUT THE INVENTION

In the following, modes for carrying out the present invention areexplained by referring to embodiments shown in the drawings.

FIG. 1 through FIG. 3 show a first embodiment of a cell/tissue cultureapparatus according to the present invention. FIG. 1 shows a wholeconfiguration of the cell/tissue culture apparatus, FIG. 2 shows aculture unit, and FIG. 3 shows a culture control device.

To this cell/tissue culture apparatus, an apparatus body 2 is provided.This apparatus body 2 is a means which forms an aseptic space 4 suitablefor a culture. A culture bath 6 is installed in an inside of thisapparatus body 2, and, as a fluid, for example, water 8 is stored in theculture bath 6. Instead of this water 8, oil and so on having properviscosity may also be used. This culture bath 6 which stores up thewater 8 may be any shape of a circular cylindrical body, a squarecylindrical body, and so on. It is desirable that the culture bath 6 isformed by a corrosion-resistant material of stainless steel and so on.

In an inside of this culture bath 6, as a culture vessel whichcultivates a cell or tissue, for example, a culture vessel 10 shown inFIG. 2 is installed. This culture vessel 10 is immersed in an interiorof the water 8, and receives a pressure namely a water pressure, whichis a physical stimulation, according to a depth at which the culturevessel 10 is immersed. The culture vessel 10 is a thing that flexiblefilms 12 and 14 which are a pressure reception member and a formingmember formed by a flexible material of rubber and so on are attached toa frame member 16. And a culture chamber 18 of a predetermined shape isformed by each of the flexible films 12 and 14. Each of the flexiblefilms 12 and 14 has properties that a heat resistance is high and thatgas and liquid are difficult to penetrate, and has a chemically stablecharacteristic that an outflow of an ingredient by dissolution is verylittle. The flexible films 12 and 14 are formed by a synthetic resinsafe and stable for a living body, for example, by a film made offluororesin. As a culture medium which becomes a scaffold of the cell ortissue, for example, a matrix 20 consisting of a bioabsorbable materialis accommodated in the culture chamber 18. The matrix 20, for example,is a thing that a bioabsorbable material of a long thread-shape isrounded so as to be tangle into one at random. Its shape is a shapesimilar to a breast of a patient, and it is a thing that reproduces ashape before an operation.

In case of this embodiment, the culture vessel 10 is fixed in the insideof the culture bath 6 by attaching the frame member 16, which maintainsthe flexible films 12 and 14, to support members 26 and 28 bythumbscrews 22 and 24 which are a fixing means. One support member 26 isprotrusively provided at a side of a bottom face of the culture bath 6,and the other support member 28 is protrusively provided at a side of aninner side wall of the culture bath 6. By this, the culture chamber 18of the culture vessel 10 which consists of the flexible films 12 and 14is set at a predetermined position in the culture bath 6.

Further, to the culture vessel 10, two tubes 30 and 32 are attached at aside of the flexible film 14, and a circulation path 36 of a culturefluid 34 is formed by these tubes 30 and 32. Therefore, for the matrix20 which is set to the inside of the culture vessel 10, the culturefluid 34 necessary for a culture is circulated and is supplied.

Further, to the culture bath 6, a means which generates oscillationand/or variation in the water 8 as a fluid is provided. In thisembodiment, a jet generation mechanism 38 is installed. The jetgeneration mechanism 38 is a thing that pressure-feeds the water 8,which is taken out of the culture bath 6 to an outside, to the inside ofthe culture bath 6 to hit on the culture vessel 10, and also generatesan oscillation for the water 8 in the culture bath 6.

In case of this embodiment, to the culture bath 6, a taking-out opening40 of the water 8 is provided at a side of its bottom face, and a returnopening 42 is formed at a side wall portion of a face side opposite tothe taking-out opening 40. Further, a circulation path 44 which makesthe water 8 circulate is formed between the taking-out opening 40 andthe return opening 42, and a pump 46 serving as a pressure-feedcirculation means of the water 8 is installed in the circulation path44. It is possible to optionally adjust the strength of a pressure-feedflow by a rotation of the pump 46.

Further, a nozzle 48 serving as a jet injection means which injects thewater 8 pressure-fed through the circulation path 44 is provided at aside of the return opening 42. In order to vary a jet generated from thenozzle 48 in vertical and horizontal directions, the nozzle 48 isattached to a vertical/horizontal operation mechanism 50. In theembodiment, a vertical direction actuator 54 serving as a verticaldirection turning means which turns the nozzle 48 in a verticaldirection with a venturi part 52 provided at an intermediate portion ofthe nozzle 48 and used as a supporting point is provided, and ahorizontal direction actuator 56 serving as a horizontal directionturning means which turns the nozzle 48 in a horizontal direction isprovided. For example, a plunger 55 of the vertical direction actuator54 is pivoted at a rear portion of the nozzle 48. In addition, amechanism turning the nozzle 48, for example, can be constituted so thatan external shape of the venturi part 52 is formed into a ball part, sothat this ball part is snapped by a rotatable support member consistingof a synthetic resin and is supported in a rotatable state by thesupport member, and so that moving force in a vertical direction or ahorizontal direction is given to an intermediate portion of the nozzle48 by using this as a supporting point.

To the vertical direction actuator 54 and the horizontal directionactuator 56, a motor is provided as a drive means, respectively. Theactuators 54 and 56 convert a rotation of each motor into a movement ofa vertical direction or a horizontal direction, and can move the nozzle48 in the vertical direction or the horizontal direction, or in thevertical and horizontal directions at the same time. That is, by turningin every direction, a directional movement over a solid angle 90° ispossible, and it is possible to uniformly inject a jet on the culturevessel 10.

Further, a means which adds a bubble flow to a jet is provided to thenozzle 48, and, in this embodiment, an air intake tube 58 is connectedto the venturi part 52 of the nozzle 48. For example, a throat 60 isformed at the venturi part 52 of an intermediate portion of the nozzle48, and the air intake tube 58 is joined to a small hole for an airintake which is formed at a constricted portion of the throat 60. Anopening end of the air intake tube 58 is installed at a positionslightly higher than the surface of water in the culture bath 6.Further, an air valve 64 which adjusts an intake of air 62 is providedto the air intake tube 58, and existence or non-existence of a bubbleflow and its amount can be adjusted by the air valve 64. The air 62which is drawn from the air intake tube 58 by a pressure-feed of a waterflow makes a bubble flow occur in the water 8, and it is possible togenerate ultrasonic waves in the water 8 by the bubble flow.

Further, to the inside of the culture bath 6, a sheathed heater 66serving as a heating means which keeps the water 8 warm is attached. Inaddition, to the taking-out opening 40 of the water 8, a temperaturesensor 68 serving as a water temperature detection means of the insideof the culture bath 6 is attached.

A detection output of the temperature sensor 68, as shown in FIG. 3, isgiven to a control unit 70 which is provided as a culture control means.The control unit 70 has a processor, a storage means, an input-outputmeans, and so on. For the control unit 70, an operation switch 72 whichorders operation of the apparatus and a stop of the operation, and aninput unit 74 serving as a means which sets a stimulation mode and so onand takes in various kinds of control outputs are provided. Further, anindication lamp 76 which performs indication of a state of operation ora stop state of the operation, and a display unit 78 serving as adisplay means which displays a state of affairs in a culture, and so onare provided. In addition, this control unit 70 generates various kindsof control outputs, and gives each control output to the pump 46, eachof the actuators 54 and 56, the heater 66 or the air valve 64.

In a composition like this, the matrix 20 of a bioabsorbable materialwhich makes a scaffold for a culture of a cell or tissue is put in theinside of the culture chamber 18, and an adipose cell is planted in thematrix 20. Because of the culture and the growth of a nerve, it isbetter to fix also NGF (Nerve Growth Factor) to the scaffold at the sametime. The culture chamber 18 in which such a matrix 20 is accommodated,namely the culture vessel 10, is immersed in the water 8 of the culturebath 6.

In the culture chamber 18, the culture fluid 34 is circulated. Alongwith this, the water 8 in the culture bath 6 is circulated through thecirculation path 44 by the pump 46, and is injected into the culturebath 6 by the nozzle 48. A water temperature in the culture bath 6 isdetected by the temperature sensor 68, a heating temperature of thesheathed heater 66 is adjusted according to the detected temperature,and the water temperature is controlled to a certain temperature.

If the pump 46 is operated, the water 8 is circulated through thecirculation path 44. Along with this, a jet is generated from the nozzle48, and the jet is hit on the culture chamber 18. A direction of the jetcan be varied by driving the two actuators 54 and 56, and it is possibleto give the jet to an optional position of a surface of the culturechamber 18. In the culture chamber 18 with which the jet collides, thatcollision part is pressed, and deformation is also caused at itscircumferential part. If an injecting direction of the jet is varied,shearing stresses of all directions are applied to the cell or tissue onthe matrix 20 of the culture chamber 18, in other wards, a massage forceis applied. For example, if the rear side of the nozzle 48 is moved inthe directions of an arrow “a” or “b” by the actuator 54, a pointed endside of the nozzle 48 moves in a vertical direction with the venturipart 52 used as a supporting point, and is to be swung in the range ofan angle θ according to magnitude of move displacement which is applied.Operation like this is also performed by the actuator 56, and the nozzle48 is to be swung in an angle range by a horizontal direction movement.

Further, if the air valve 64 is opened, since the venturi part 52 isunder a state of a negative pressure due to the occurrence of a jet bythe pump 46, the air 62 is sucked into the jet. By this, the air 62becomes minute bubbles, mixes with the jet, and is injected into thewater 8 of the culture bath 6 from the nozzle 48. The minute bubblesgenerate ultrasonic waves, give a vibrational stimulation to the cell ortissue on the matrix 20, and produce a good effect.

Like this, an optimum pressure is maintained by a pressure by-pass valvenot shown in the drawings while a shearing stress due to a jet of thewater 8 is being given to a cell or tissue on the matrix 20, and, underthis state, it is possible to make the culture fluid 34 circulate in theculture chamber 18 and to cultivate the cell or tissue. Further, byvarying the velocity of a jet by control of a rotating speed of the pump46, and so on, it is possible to give an optimum physical stimulation tothe cell or tissue on the matrix 20. In this case, strength of the jetgiven to the cell or tissue, for example, is controlled by a programwhich varies a rotating speed of the pump 46 so as to make an optimumstress with the lapse of a culture time. Further, by adjustment of anamount or water level of the water 8, it is also possible to adjust ahydrostatic pressure given to a culture part. As a result, it ispossible to give physical stimulation of two kinds of the hydrostaticpressure and the shearing stress.

In this embodiment, the culture vessel 10 is fixed to the culture bath6. However, a composition able to vary a depth of immersion to the water8 can be given, and, if the magnitude of a water pressure applied to theculture vessel 10 namely the culture chamber 18 is varied, it ispossible to give an optimum physical stimulation due to variation in awater pressure to the cell or tissue on the matrix 20. Further, in casein which the culture vessel 10 is fixed to the culture bath 6, byvarying an amount of the water 8, likewise it is possible to give aphysical stimulation due to a water pressure. Furthermore, by using asterilizing fluid for the water 8 in the culture bath 6, and byperforming the improvement of bacteria-proof of the aseptic space 4 andthe culture bath 6, it is possible to give higher contamination controlof the prevention of admission of bacteria to the culture chamber 18,and so on.

Furthermore, according to this cell/tissue culture apparatus, it ispossible to give a shearing stress to the whole of a tissue under aculture from a random direction. That is, by giving the shearing stressto the whole of the tissue from a random direction, in other words, bygiving a massage force, it is possible to cultivate a tissue with a lotof gapes, namely, a tender adipose tissue. In the adipose tissue havinga lot of gapes, flexibility is high, and, after transplanting to aliving body, the creation of a blood vessel and a nerve cell becomeseasy and can be made to promote. Therefore, if an adipose tissue whichis cultivated by such a culture apparatus is used, it is possible torestore a breast serving as a tissue body having blood and nervesequivalently to a human body. Further, such a culture apparatus can alsobe used for a culture of an adipose tissue of a pancreas (Langerhans'islet), and can contribute to the medical treatment of diabetes andpancreas cancer.

An example of operation of a control program of this cell/tissue cultureapparatus is explained by referring to flow charts in FIG. 4 throughFIG. 6. In FIG. 4 through FIG. 6, reference letters “a”, “b”, “c”, “d”,“e”, “f” and “g” show connectors among the flow charts.

A step S1 shows an input process of an operation program by means of theinput unit 74. After a predetermined operation program is set, if theoperation switch 72 is closed at a step S2, the indication lamp 76 islighted up at a step S3, and indication representative of a state underthe operation is performed. Further, at a step S4, on the basis of acomparison between a water temperature taken from the temperature sensor68 and a set temperature, a heating temperature of the sheathed heater66 is adjusted, and a temperature of the water 8 is controlled to acertain value.

The pump 46 is driven by the closing of the operation switch 72, and, ata step S5, the pump 46 is driven at jet strength J₁ consisting of lowstrength at the time of a start of the culture. At the following stepS6, the air valve 64 is opened, and a motor of each of the actuators 54and 56 is driven. As a result, a jet accompanied with a bubble flow isemitted in the direction of the culture chamber 18 from the nozzle 48,and a shearing stress due to the jet is given to the matrix 20 under theculture. By driving each of the actuators 54 and 56, the jet from thenozzle 48 is uniformly hit on a surface of the culture chamber 18.

Further, at a step S7, whether or not abnormality exists in the pump 46and motors of the actuators 54 and 56 is decided. In case in whichabnormality does not exist, processing proceeds to a step S8, and theoperation of the pump 46 is continued by a time T_(J1) (a jet time).After the lapse of the time T_(J1), the processing proceeds to a stepS9, and the pump 46 is stopped.

At a step S10, temperature control of the water 8 similar to the step S4is executed, and, at a step S11, an abnormality decision similar to thestep S7 is performed. In case in which abnormality does not exist, theprocessing proceeds to a step S12, and waiting for the lapse of a timeT_(S1)(a jet stop time) is performed. Then, the processing proceeds to astep S13, and waiting for the lapse of a time T₁ (a culture continuationtime of a first stage) is performed. After that, the processing proceedsto a step S14, and temperature control similar to the step S4 isexecuted.

At a step S15, the pump 46 is driven at jet strength J₂ consisting ofhigh strength. At a step S16, whether or not abnormality exists in thepump 46 and the motors of the actuators 54 and 56 is decided again. Incase in which abnormality does not exist, the processing proceeds to astep S17, and the operation of the pump 46 is continued by a time T_(J2)(a jet time). After the lapse of the time T_(J2), the processingproceeds to a step S18, and the pump 46 is stopped. Then, the processingproceeds to a step S19, and temperature control similar to the step S4is performed.

Further, at a step S20, whether or not abnormality exists in the pump 46and the motors of the actuators 54 and 56 is decided again. In case inwhich abnormality does not exist, the processing proceeds to a step S21,and waiting for the lapse of a time T_(S2) (a jet stop time) isperformed. After the lapse of the time T_(S2), the processing proceedsto a step S22, and waiting for the lapse of a time T₂ (a culturecontinuation time of a second stage) is performed. Then, the processingproceeds to a step S23, and temperature control similar to the step S4is executed.

Further, at a step S24, whether or not abnormality exists in the pump 46and the motors of the actuators 54 and 56 is decided again. In case inwhich abnormality does not exist, the processing proceeds to a step S25,and the end indication of a culture is performed at the time of acompletion of the culture. Then, at a step S26, whether or not theoperation switch 72 is in OFF is decided. In case of OFF, a stop of thepump 46, a close of the air valve 64 and a stop of each of the actuators54 and 56 are performed, and, at a step S28, a cancellation of theoperation indication and a cancellation of the end indication areperformed.

Furthermore, in case in which a decision that abnormality exists isgiven at the steps S7, S11, S16, S20 and S24, the processing proceeds toa step S29, and the operation of the pump 46 and the actuators 54 and 56is stopped. Further, at a step S30, a warning display is performed onthe display unit 78 as a notification of abnormality.

Control operation like this can be represented by a timing chart shownin FIG. 7, for example. In FIG. 7, a reference letter (A) shows theoperation indication, and a reference letter (B) shows the endindication. A reference letter (C) shows the operation time and the stoptime of the pump 46, and strength and weakness (J₁, J₂) of a water flowthereof. A reference letter (D) shows the movement in a verticaldirection of the actuator 54, and a reference letter (E) shows themovement in a horizontal direction of the actuator 56. A referenceletter (F) shows the opening and closing of the air valve 64, areference letter (G) shows a power-supplying interval and apower-supplying stop interval of the sheathed heater 66, and a referenceletter (H) shows the lapse of a detected temperature of the temperaturesensor 68. As is clear from the control operation shown in FIG. 7, it ispossible to vary as shown in the jet strength J₁, J₂ by an adjustment ofa rotating speed of the pump 46.

Next, FIG. 8 shows a second embodiment of a cell/tissue cultureapparatus according to the present invention.

The cell/tissue culture apparatus of this embodiment, similarly to thefirst embodiment, immerses a culture vessel 10 in water 8 of a culturebath 6, and cultivates a cell or tissue on a matrix 20 in a culturechamber 18. However, this apparatus applies a shearing stress to thecell or tissue on the matrix 20 by waves serving as an oscillation ofthe water 8 instead of the physical stimulation by means of the jet ofthe water 8 in the first embodiment. Thereby, a physical stimulation isgiven to the cell or tissue.

In this embodiment, the culture vessel 10 namely the culture chamber 18is provided in the water 8 of the culture bath 6 on the float. Amaterial, a composition and a shape of the culture chamber 18, and thematrix 20 of a scaffold and soon for a culture of its inside are similarto the first embodiment. In this case, a wave generating plate 80serving as an oscillation generating means is provided at a side wallpart of the culture bath 6. To this wave generating plate 80, arotational force which is generated by a driving means of a motor 82 andso on is converted into an angle variation 0 by a link mechanism 84 andis given. Waves (standing waves) are generated in the water 8 of theinside of the culture bath 6 by means of the angle variation θ of thewave generating plate 80. As a result, it is possible to apply aphysical stimulation due to the waves to the cell or tissue in theculture chamber 18.

Instead of a mechanical means of the wave generating plate 80 and themotor 82, the oscillation generating means may also be constituted sothat the wave generating plate 80 is vibrated by a mechanism whichconverts an electric signal into a mechanical vibration, for example, bya solenoid such as a voice coil of a speaker. In this case, it ispossible to generate a vibration by a frequency signal suitable for acell or tissue.

Further, applying such a vibration to the water 8 in the culture bath 6,the culture chamber 18 is to receive various stresses of a bendingstress, a shearing stress, a compressive stress and so on by the waves,and the cell or tissue on the matrix 20 of its inside receives mainly ashearing stress from various directions. Like this, the apparatuscultivates a cell or tissue while giving a physical stimulation by meansof the waves.

In this case, varying the strength or frequency of the waves, forexample, by a manner which controls a revolution speed or a revolutionstroke of the motor 82, or by a manner which varies a current value, afrequency and/or a power-supplying interval given to the voice coil, itis possible to give a stimulation suitable for a cell or tissue to becultivated. In addition, the strength or frequency of a physicalstimulation given to a cell or tissue can be controlled according to aprogram which makes the strength or the frequency vary so as to make anoptimum stress with the lapse of a culture time, and an effect similarto the first embodiment is obtained.

According to the cell/tissue culture apparatuses descried above, it ispossible to give an optional physical stimulation of a shearing stressand so on to a cell or tissue under a culture by an oscillation of apressure or a fluid. Further, it is possible to cultivate a tissue bodyaccording to a shape of a desired portion of a human body, and it isalso possible to realize flexibility and toughness which that portionhas.

The configurations, the actions and the effects as the modes forcarrying out the present invention are described by referring to theembodiments shown in the drawings. However, the present invention is notlimited to the modes for carrying out the present invention and theembodiments mentioned above. The present invention includes allconfigurations which can be predicted or conjectured by a person skilledin the art, namely, various kinds of compositions, modified examples,and so on, which are conjectured from the objects of the invention, themodes for carrying out the invention, and the embodiments of theinvention.

INDUSTRIAL APPLICABILITY

As descried above, the cell/tissue culture apparatus according to thepresent invention gives an optimum physical stimulation of a shearingstress and so on to a cell or tissue under a culture, can be utilizedfor a culture of a tissue body according to a shape of a desired portionof a human body, and realizes flexibility and toughness which thatportion has. Because of this, the present invention is very useful for aculture of a cell or tissue.

1. A cell/tissue culture apparatus comprising: a culture chamber beingformed by a flexible material that accommodates a cell or tissue to becultivated and also makes a culture fluid circulate; and said culturechamber being immersed in a fluid.
 2. The cell/tissue culture apparatusof claim 1, wherein, by making said fluid oscillate, a physicalstimulation is given to said cell or tissue ins aid culture chamber. 3.The cell/tissue culture apparatus of claim 1, wherein said culturechamber provides a space part of a shape according to a shape of cellsor tissue to be cultivated.
 4. The cell/tissue culture apparatus ofclaim 2, wherein said oscillation of the fluid is due to a jet of afluid.
 5. The cell/tissue culture apparatus of claim 2, wherein saidoscillation of the fluid is due to wave motion.
 6. The cell/tissueculture apparatus of claim 1, wherein said physical stimulation is ashearing stress.
 7. The cell/tissue culture apparatus of claim 1,wherein said physical stimulation is given intermittently orcontinuously.